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PMID: 3018532 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Analysis of an activatable promoter: sequences in the simian virus 40 late promoter required for T-antigen-mediated trans activation.

Molecular and cellular biology ·Vol. 5 ·No. 8 ·1985-08-00 ·Pages 1859-69

Keller JM, Alwine JC

Abstract

The late promoter of simian virus 40 (SV40) is activated in trans by the viral early gene product, T antigen. We inserted the wild-type late-promoter region, and deletion mutants of it, into chloramphenicol acetyltransferase transient expression vectors to identify promoter sequences which are active in the presence of T antigen. We defined two promoter activities. One activity was mediated by a promoter element within simian virus 40 nucleotides 200 to 270. The activity of this element was detectable only in the presence of an intact, functioning origin of replication and accounted for 25 to 35% of the wild-type late-promoter activity in the presence of T antigen. The other activity was mediated by an element located within a 33-base-pair sequence (simian virus nucleotides 168 to 200) which spans the junction of the 72-base-pair repeats. This element functioned in the absence of both the origin of replication and the T-antigen-binding sites and appeared to be responsible for trans-activated gene expression. When inserted into an essentially promoterless plasmid, the 33-base-pair element functioned in an orientation-dependent manner. Under wild-type conditions in the presence of T antigen, the activity of this element accounted for 65 to 75% of the late-promoter activity. The roles of the 33-base-pair element and T antigen in trans-activation are discussed.

MeSH Terms
Acetyltransferases/genetics Animals Antigens, Polyomavirus Transforming Antigens, Viral, Tumor/genetics Cell Line Chloramphenicol O-Acetyltransferase Chlorocebus aethiops Chromosome Deletion DNA Replication DNA Restriction Enzymes Genes Genes, Viral Kidney Oncogene Proteins, Viral/genetics Plasmids Promoter Regions, Genetic Protein Kinases/genetics Simian virus 40/genetics Transfection Virus Activation
Chemicals
Antigens, Polyomavirus Transforming Antigens, Viral, Tumor Oncogene Proteins, Viral Acetyltransferases Chloramphenicol O-Acetyltransferase Protein Kinases DNA Restriction Enzymes
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Keller J M
Alwine J C
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.5.8.1859-1869.1985
Published
1985-08-00
Pages
1859-69
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC366901
Subset
IM
Grants
NCI NIH HHS · CA28379 · United States
NCI NIH HHS · CA33656 · United States
NCRR NIH HHS · S07-RR-05145-23 · United States
External Links
PubMed source DOI: 10.1128/mcb.5.8.1859-1869.1985 PMC Full Text View journal details
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